This project aims to demonstrate the feasibility of an in vivo diagnostic platform for recognizing indicators of tumor-microenvironment, unambiguously, in biological fluids, tissue phantoms and small animal imaging. Specific project aims are to synthesize and evaluate cancer-specific, thermoresponsive, nanoparticle-based, near-infrared (NIR) fluorescent contrast agents (ThermoDots) for optical imaging of tumors. ThermoDots will increase the payload and spatial resolution of the delivered contrast agent from the combined effect of: (1) enhanced signal of NIR fluorophores due to temperature modulation (TM) by high-intensity focused ultrasound (HIFU) agitation, and (2) targeted binding of prostate cancer-specific moieties conjugated onto the particle surface. Phase I tasks include benchmarking experiments to validate the design in relevant physiological media, tissue phantoms, cell studies and preliminary in vivo imaging using an orthotopic cancer model (PC-3M) in nude mice. The recognition relevance of ThermoDots will undergo second-stage validation in Phase II in the extended animal model studies using temperature modulated fluorescence tomography (TM-FT) guided HIFU small animal imaging. The project team includes a university collaborator with extensive experience in the tumor imaging area.